MiaA (Rv2727c) mediated tRNA isopentenylation of Mycobacterium tuberculosis H37Rv

Document Type : Original article

Authors

1 School of Biotechnology, Gautam Buddha University, Gautam Budh Nagar, Greater Noida, Uttar Pradesh, India

2 School of Sciences, Indira Gandhi National Open University, Maidan Garhi, New Delhi, India

Abstract

tRNA modifications play a significant role in the structural stability as well as translational fidelity in all organisms from bacteria to humans. They also play a major role in bacterial physiology by regulating translation in response to various environmental stresses. Modifications coming at the anticodon-stem loop (ASL) are particularly important as they stabilize codon-anticodon interactions, ensuring accuracy and speed in decoding mRNAs  Addition of isopentenyl group (i6A) at A37 position by tRNA isopentenyltransferase (MiaA) is a well conserved modification from bacteria to human. We studied M. tuberculosis MiaA from strain H37Rv and identified the target tRNAs for this modification based on the A36A37A38 motif. i6A modification of target tRNAs tRNALeuCAA, tRNAPheGAA, tRNATrpCCA and tRNASerCGA were further confirmed by isopentenyltransferase assay providing the substrate DMAPP and recombinant MiaA enzyme.

Keywords

References

  1. WHO. Global tuberculosis reports 2020, Global Tuberculosis Reports. (2020). https://apps.who.int/iris/bitstream/handle/10665/336069/9789240013131-eng.pdf.
  2. Delogu G, Sali M, Fadda G. The biology of mycobacterium tuberculosis infection. Mediterr J Hematol Infect Dis 2013;5:e2013070.
  3. Sasindran SJ, Torrelles JB. Mycobacterium tuberculosis infection and inflammation: what is beneficial for the host and for the bacterium? Front Microbiol 2011;2:2.
  4. Wang X, He Ch. Dynamic RNA modifications in posttranscriptional regulation. Mol Cell 2014;56:5-12.
  5. Chionh YH, McBee M, Babu RI, Hia F, Lin W, Zhao W, Cao J, Dziergowska A, Malkiewicz A, Begley TJ, Alonso S, Dedon PC. tRNA-mediated codon-biased translation in mycobacterial hypoxic persistence. Nat Commun 2016;7:13302.
  6. Schweizer U, Bohleber S, Fradejas-Villar N. The modified base isopentenyladenosine and its derivatives in tRNA, RNA Biol 2017; 1197-1208.
  7. Lorenz C, Lünse CE, Mörl M. tRNA Modifications: impact on structure and thermal adaptation. Biomolecules 2017;7:35.
  8. Thompson KM, Gottesman S. The MiaA tRNA modification enzyme is necessary for robust RpoS expression in escherichia coli. J Bacteriol 2014;196:754-761.
  9. Durand JM, Björk GR, Kuwae A, Yoshikawa M, Sasakawa C. The modified nucleoside 2-methylthio-N6-isopentenyladenosine in tRNA of Shigella flexneri is required for expression of virulence genes. J Bacteriol 1997;179:5777-5782.
  10. Goude R, Parish T. Electroporation of mycobacteria. in: T. Parish, A.C. Brown (Eds.), Mycobacteria Protocols: Second Edition, Humana Press, Totowa, NJ. Methods Mol Biol 2009;465:203-215.
  11. Kaur P, Rausch M, Malakar B, Watson U,  Damle NP, Chawla Y, Srinivasan S, Sharma K, Schneider T, Jhingan GD, Saini D, Mohanty D, Grein F, Nandicoori VK. LipidII interaction with specific residues of Mycobacterium tuberculosis PknB extracytoplasmic domain governs its optimal activation. Nat Commun 2019;10:1231.
  12. Chambers AE, Richardson AE, Read DF, Waller TJ, Bernstein DA, Smaldino PJ. An in vitro assay to detect tRNA-isopentenyl transferase activity. J Vis Exp 2018;8:58100.
  13. Soderberg  T,  Poulter  CD, Escherichia coli dimethylallyl diphosphate:tRNA dimethylallyl-transferase: essential elements for recognition of tRNA substrates within the anticodon stem-loop. Biochemistry 2000;39:6546-6553.
  14. Heuston S, Begley M, Gahan CGM, Hill C. Isoprenoid biosynthesis in bacterial pathogens. Microbiology 2012;158:1389-1401.
  15. Pierrel F, Douki T, Fontecave M, Atta M. MiaB protein is a bifunctional radical-S-adenosylmethionine enzyme involved in thiolation and methylation of tRNA. J Biol Chem. 2004;279:47555-47563.
  16. Wei FY, Zhou B, Suzuki T, Miyata K, Ujihara Y, Horiguchi H, Takahashi N, Xie P, Michiue H, Fujimura A, Kaitsuka T, Matsui H, Koga Y, Mohri S, Suzuki T, Oike Y, Tomizawa K. Cdk5rap1-mediated 2-methylthio modification of mitochondrial tRNAs governs protein translation and contributes to myopathy in mice and humans, Cell Metab. 2015;21:428-442.
  17. Lamichhane TN, Arimbasseri AG, Rijal K, Iben JR, Wei FY, Tomizawa K, Maraia RJ. Lack of tRNA-i6A modification causes mitochondrial-like metabolic deficiency in S. pombe by limiting activity of cytosolic tRNATyr, not mito-tRNA. RNA 2016;22:583-596.
  18. Yarham JW, Lamichhane TN, Pyle A, Mattijssen S, Baruffini E, Bruni F, Donnini C, Vassilev A, He L, Blakely EL, Griffin H, Santibanez-Koref M, Bindoff LA, Ferrero I, Chinnery PF, McFarland R, Maraia RJ, Taylor RW. Defective i6A37 modification of mitochondrial and cytosolic tRNAs results from pathogenic mutations in TRIT1 and its substrate tRNA. PLoS Genet 2014;10:e1004424.
  19. Aubee JI, Olu M, Thompson KM. The i6A37 tRNA modification is essential for proper decoding of UUX-Leucine codons during rpoS and iraP translation. RNA 2016;22:729-742.
  20. Blango MG, Fleming BA, Kincannon WM, Tran A, AJ Lewis, Russell CW, Zhou Q, Baird LM, Brannon JR, Beebout CJ, Bandarian V, Hadjifraniskou M, Howard MT, Mulvey MA. Balanced input from the tRNA prenyltransferase MiaA controls the stress resistance and virulence potential of extraintestinal pathogenic escherichia coli. BioRxiv 2021;02:429414.

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